人脐静脉内皮细胞的培养

Culture of the Endothelial Cells Derived from the Human Umbilical Cord Vein in Vitro

目的 :培养的内皮细胞为基础和临床研究提供体外模型 ,并为进一步研究内皮细胞对造血的影响提供实验材料。方法 :经胶原酶消化获得人脐静脉内皮细胞 ,用RPMI 164 0加 2 0 %的胎牛血清进行培养。结果 :成簇状的内皮细胞最初形成于培养皿 (或培养瓶 )的底层 ,继而细胞迅速生长 ,融合成片 ,约 7天细胞呈单层多角形镶嵌排列。透射电镜观察 ,原代培养细胞的胞质内含内皮细胞特有的细胞器 (WPBs)。兔抗人Ⅷ因子相关抗原 (ⅧR∶Ag)免疫组织化学染色实验证实 ,培养的内皮细胞含有ⅧR∶Ag。 结论 :本实验的各项结果从形态 ,免疫组化及透射电镜的观察均符合内皮细胞的特征 ,因此确信所培养的内皮细胞可提供作进一步深入的细胞生物学及对造血影响的研究。

s Objective:[WT5”BZ]Cultured endothelial cells provide a useful in vitro model for the study on the basic and clinical medicine and the experimental materials for the further study of the endothelial cells to hematopoiesis.Methods:Human endothelial cells freshly obtained from human umbilical cords by collagenase digestion of the interior of the umbilical vein were cultrued using RPMI 1640 supplemented with 20% fetal calf serum.Results:Small clusters of the cells inilially spread on the bottom of the dishs or bottles,then coalesced and grew to form confluent monolayers of polygonal cells about 7 day.Under transmission electron microscope,cells in primary culturs were identified as endothelium by the presence of the specific cytoplasmic organelle(Weibel Palade bodies).The immunohistochemical staining of the rabbit anti factor Ⅷ related antigen(ⅧR∶Ag)showed the cultured endothelial cells containing ⅧR∶Ag.Conclusion:The cells,identified as endothelial cells according to morphologic,immunohistochemical and transmission electron microscopical characteristics,could be useful for further studying cell biology and in the research of their influence to hematopoiesis.